【摘要】 旨在月季中建立高效、快速的VIGS实验技术体系,以期能够改进VIGS技术体系在月季等木本植物表达效率低等缺陷。采用RT-PCR方法进行月季RhPDS基因片段的克隆,在以pTRV2为原载体,采用双酶切的方法构建了pTRV2-RhPDS重组载体。经双酶切及测序验证后,通过电击法转化农杆菌GV3101,并分别采用高压喷枪法对月季植株进行侵染、采用真空渗透法对扦插苗以及组培苗进行侵染。采用qRT-PCR方法分析RhPDS基因的表达。结果显示:构建的重组载体pTRV2-RhPDS,经双酶切验证,显示条带单一清晰,符合预期目标。在侵染方法和材料上,采用真空渗透法侵染组培苗是最为快速、高效的方式。该试验获得了较为高效、快速VIGS技术体系,提高了VIGS技术体系在月季上表达效率。更多还原

【Abstract】 This study established an efficient and fast VIGS experimental system in rosa hybrida to improve the expression efficiency in rosa hybrida and other woody plants by VIGS.The coding sequence of RhPDS was amplified using RT-PCR method.pTRV2 was used as the original vector,and pTRV2-RhPDS vectors were constructed by double digestion.The test result was validated by double digestion and sequencing,and then the VIGS expression vectors were transformed into Agrobacterium strain GV3101 by eletroporation.The whole plant was sprayed by a portable air compressor.The vacuum infiltration method was used for rose cuttings and tissue culture.The expression level of RhPDS was detected by RT-qPCR.The results showed that the recombinant vector pTRV2-RhPDS,verified by double enzyme digestion,had a single and clear band,which met the target.Considering the methods and materials,vacuum osmosis was the most rapid and efficient way to infect tissue culture seedling.In this research,we obtained an optimized VIGS technical system and improved the expression efficiency of VIGS in rosa hybrida.更多还原